Adenovirus Test Methods and Reference Publications Extraction Reference ...

Adenovirus Test Methods and Reference Publications

Extraction

Prior to adenovirus testing, all sample types including stool, respiratory specimens and whole blood

should be extracted. Laboratories have used the easyMAG or eMAG instruments with good results.

Alternative extraction platforms including the MagNAPure 96 have been validated for whole blood for

other viruses such as Zika. Additional extraction platforms and specimen types may be validated and

linked to the molecular assays listed below.

Reference for real-time PCR Adenovirus methods

Pan adenovirus methods:

a. Heim A, Ebnet C, Harste G, Pring-Akerblom P. Rapid and quantitative detection of human

adenovirus DNA by real-time PCR. J Med Virol. 2003;70(2):228-39.

b. Damen M, Minnaar R, Glasius P, van der Ham A, Koen G, Wertheim P, Beld M. Real-time PCR with

an internal control for detection of all known human adenovirus serotypes. J Clin Microbiol.

2008;46(12):3997-4003.

Detection of Adenovirus subgenus F (types 40 and 41) :

a. Lyman WH, Walsh JF, Kotch JB, Weber DJ, Gunn E, Vinj¨¦ J. Prospective study of etiologic agents of

acute gastroenteritis outbreaks in child care centers. J Pediatr. 2009;154(2):253-7.

b. Mori K, Hayashi Y, Akiba T, Nagano M, Tanaka T, Hosaka M, Nakama A, Kai A, Saito K, Shirasawa

H. Multiplex real-time PCR assays for the detection of group C rotavirus, astrovirus, and Subgenus

F adenovirus in stool specimens. J Virol Methods. 2013;191(2):141-7.

Pan adenovirus & HAdV-40 and -41 type-specific detection:

a. Jothikumar N, Cromeans TL, Hill VR, Lu X, Sobsey MD, Erdman DD. Quantitative real-time PCR

assays for detection of human adenoviruses and identification of serotypes 40 and 41. Appl

Environ Microbiol. 2005;71(6):3131-6.

References for Adenovirus typing methods based on Sanger Sequencing

Typing based on the six hypervariable regions (HVR 1-6) of the hexon gene:

a. Lu X, Erdman DD. Molecular typing of human adenoviruses by PCR and sequencing of a partial

region of the hexon gene. Arch Virol. 2006;151(8):1587-602.

b. Okada M, Ogawa T, Kubonoya H, Yoshizumi H, Shinozaki K. Detection and sequence-based typing

of human adenoviruses using sensitive universal primer sets for the hexon gene. Arch Virol.

2007;152(1):1-9.

Typing based on HVRs 1-6 of the hexon gene and the fiber gene:

a. McCarthy T, Lebeck MG, Capuano AW, Schnurr DP, Gray GC. Molecular typing of clinical

adenovirus specimens by an algorithm which permits detection of adenovirus coinfections and

intermediate adenovirus strains. J Clin Virol. 2009;46(1):80-4.

Typing based on hypervariable region 7 (HVR7) of the hexon gene:

a. Casas I, Avellon A, Mosquera M, Jabado O, Echevarria JE, Campos RH, Rewers M, Perez-Bre?a P,

Lipkin WI, Palacios G. Molecular identification of adenoviruses in clinical samples by analyzing a

partial hexon genomic region. J Clin Microbiol. 2005;43(12):6176-82.

Culture

Adenovirus culture

a. Cromeans TL, Lu X, Erdman DD, Humphrey CD, Hill VR. Development of plaque assays for

adenoviruses 40 and 41. J Virol Methods. 2008 Jul;151(1):140-5. doi:

10.1016/j.jviromet.2008.03.007. Epub 2008 Apr 25. PMID: 18440077.

For general adenovirus culture on respiratory and stool samples, A-549 cells can be used. Other cell lines

such as HEP-2, CaCO2 or HEK-293T may be used for enhanced culture, which can be necessary when

culturing adenovirus from blood. Adenovirus culture is often more successful if the initial inoculation is

onto monolayers in shell vials or wells in a plate. For best results, centrifugation enhancement followed

by passed/blind passing to a second culture may be necessary, depending on the extent of CPE.

ATCC CRL for Heck-293T for adenovirus 41 isolation

NGS

Metagenomics

a. Sofia Morfopoulou, Vincent Plagnol, Bayesian mixture analysis for metagenomic community

profiling, Bioinformatics, Volume 31, Issue 18, 15 September 2015, Pages 2930¨C

2938,

GenBank Accession No. for HAdV-41 sequences

MT790999.1

MW567963.1

MW686856.1

MW686855.1

MW686854.1

MW567966.1

MW567965.1

MW567964.1

MG925782.1

AB610519.1

D13781.1

DQ315364.2

AB330122.1

X51783.1

KY316163.1

KY316162.1

KY316160.1

MH465394.1

AB610523.1

HM565136.3

MT790998.1

KF303071.1

KF303070.1

KF303069.1

MW686853.1

KX868523.2

KY316164.1

AB728839.1

MW567962.1

MW686857.1

KY316161.1

MG925783.1

HQ005289.1

AB610527.1

AB610526.1

AB610525.1

AB610524.1

AB610522.1

AB610521.1

AB610520.1

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